SEPARATION OF ESCHERICHIA-COLI RNA-POLYMERASE SIGMA-70 HOLOENZYME FROM CORE ENZYME ON HEPARIN-SEPHAROSE COLUMNS

被引：5

作者：

WELLINGTON, SR ^{[1
]}

SPIEGELMAN, GB ^{[1
]}

机构：

[1] UNIV BRITISH COLUMBIA,DEPT MICROBIOL,VANCOUVER V6T 1W5,BC,CANADA

来源：

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS | 1991年 / 179卷 / 02期

基金：

加拿大自然科学与工程研究理事会;

关键词：

D O I：

10.1016/0006-291X(91)91934-5

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

A method is described for the rapid purification of DNA-dependent RNA polymerase sigma-70 holoenzyme from Escherichia coli. The essential step in this protocol involves the differential elution of sigma-70 holoenzyme from core polymerase on a heparin-Sepharose column. Using a linear gradient of KCl, holoenzyme was found to elute at 0.25M whereas core polymerase eluted at 0.35 M. From 20 g of cells, up to 1 mg of RNA polymerase holoenzyme could be isolated in two days. The preparations were greater than 95% pure with respect to protein, and saturated with the sigma subunit. © 1991.

引用

页码：1107 / 1114

页数：8